Galectin-3 plasmapheresis therapy

ABSTRACT

A system and method for the practice of apheresis employs modules in the system which can be selected for a particular patient to treat particular situations or combinations of difficulties. In one example, Gal-3 mediates a large number of body reactions, and is an effective protector of tumor microenvironments and the like, as well inflammation driver. Removal of Gal-3 may make antic-cancer treatments, like photopheresis and TNF administration more effective. Separate modules, such as one for photopheresis and one for TNF receptor removal, may be combined with a module for the reduction of Gal-3, to render the combination of treatments each more effective than if administered alone.

PRIORITY DATA AND INCORPORATION BY REFERENCE

This application claims benefit of priority to U.S. Provisional PatentApplication No. 62/139,026 filed Mar. 27, 2015 and PCT PatentApplication Serial No: PCT/US14/38694 which are incorporated herein byreference in their entirety.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to apparatus and methods employed toselectively treat agents in a mammal's blood outside of its body. Inconjunction with such treatment, generally referred to as apheresis,agents can be added to improve or effect treatment of various diseaseconditions.

Statement of Related Cases

This application is related in character and function to U.S. Pat. No.8,764,695 and U.S. patent application Ser. No. 14/141,509 filed Dec. 27,2013, both to the same inventive entity of this application. Those twocases, the disclosures of which are incorporated herein-by-reference,are focused on the use of apheresis to selectively remove galectin-3from a mammalian patient's bloodstream, to effect treatment of at leastone of inflammation, fibrosis and cell proliferation. These patentpublications focus on the use of selective agents, as opposed to generalfilter membranes and the like, to extract active or circulatinggalectin-3 from a patient's blood supply.

This application is also related to Patent Cooperation TreatyApplication PCT/US14/38694 filed May 20, 2014. The entire disclosure ofthe related application is incorporated by reference herein as well. Thelater-filed case is directed to a apheresis device optimized toselectively remove galectin-3 and/or other blood components to bettereffect treatment of these patients in a variety of fields. The devicefeatures removable and replaceable “cartridges” or similar filters thatallow for some optimization of the individual's treatment, therebyimproving results because the removal and addition of contents is moreclosely tailored to the individual's needs.

SUMMARY OF THE INVENTION

Extended development of the apheresis treatment that originally focusedon the removal of galectin-3 has revealed the value of apheresis in thetreatment of a variety of mammalian situations that are not necessarilylimited to galectin-3 removal, or selective removal of galectin-3 atall. Indeed, galectins make up a family of compounds which mediate awide variety of biological functions, including everything frominflammation to uterine implantation and multiple aspects ofhomeostasis. Galectin-1, galectin-3, galectin-9 are among those mostwidely focused on, but all of the galectins in a mammal may be targetsfor treatment. In this application, a target galectin, such asgalectin-3, may typically be removed in the treatment that an agentinhibited or medicated by galectin-3 is addressed as unrelatedtechnologies, such as 3D printing, cancer treatment strategies and moreclosely aligned technologies such as personalized medicine have grownand developed, the inventor has recognized that the synthesis ofapheresis with these other technologies opens up new doors and avenuesto treatment that have not been fully explored.

Apheresis is conventionally defined as the removal of blood from adonor, typically a mammal, and more specifically preferably a human, acompanion animal or a commercial animal, which may include separation ofblood components, followed by return of most of those components to thepatient by retransfusing the whole blood back to the patient. In mostapheresis devices and treatments, blood cells are separated from plasmainitially, and the plasma is subsequently treated to at least remove ablood component before return to the individual. Blood cells, whichtypically entrain platelets and related blood components, may also betreated (or collected for specific purposes such as to provide donormaterial). By removing the blood and plasma from the body, treatment ofa wide variety of conditions and disorders becomes simplified, withouthaving to deal, at least initially, with natural responses and sideeffects, such as cytokine-triggered inflammation responses. Thus, anentire new range of therapies may be offered that would be otherwisefrustrated or limited by natural body defenses if administered to thepatient in vivo, including blocking agents, cytokines, antibodies andthe like. Thus, the treatment method contemplated herein is ex vivo,blood is removed, and it, or if preferred components thereof, aretreated, to remove compounds, to add agents, and to specifically modifycompounds or modulate aspects of the internal environment, or to collectspecific cell types, before returning the altered fluids to the patientto effect treatments made possible by those alterations. Both thetreatment methods, and the devices to effect those treatments, arecontemplated herein.

This invention recognizes that ultimately, a positive treatmenteffective in the amelioration of one disease state in one patient, e.g.,a cancer therapy, may be best deployed to treat the same disease in adifferent patient slightly differently, to account for differences inthe two patients that may be due to age, race, genetic and epigeneticviability, co-morbidities such as obesity, diabetes, lung function andcirculatory condition, disease stage, concomitant therapies, etc. Wherefor instance treatment is preventive rather than therapeutic, such as anattempt to remove specific potential cancerous cells or the like,genetic or other determinants may vary individual to individual. Itwould be of value to develop a general treatment that could be optimizedor personalized for each individual, without the substantial costcurrently associated with current applications of “personalizedmedicine.” By taking advantage of the localization of the blood outsideof the patient's body provided for by apheresis, a whole host ofpatient-specific modifications can be practiced. Many of thesestrategies are enhanced by being conducted ex vivo, so that bodychemistry issues can be dealt with more selectively, or avoided.

Broadly stated, the invention calls for the adoption of an apheresisstrategy specific to the individual. This includes the opportunity todeploy apheresis wherein apheresis may be used as a control device tomake adjustments outside of the body instead of inside the body, wherethe inflammatory response is much more difficult to control as in thecase of the selective removal of galectin-3 (“Gal-3”), as described inthe aforementioned patents and applications. This could involvepre-treatment diagnostic to determine the serum level of Gal-3 in thepatient, so as to identify the number of filters or cartridges requiredto remove the necessary amount. (“Gal-3” as used herein refers tocirculating Gal-3 able to bind to receptors and molecules. Gal-3 existsin at least two, possibly three, forms, one of which is pentomeric andone which may be monomeric, but a dimer is also possible. It may also bebound by several polysaccharides. Gal-3 as used herein refers to allthese forms.) It might further involve therapeutic or preventivediagnosis to prescribe agents or additives best added to the patient'sbloodstream before return to the body, for instance, cytokines oraltered cells, pharmaceuticals or natural agents, etc. for, e.g., aparticular cancer therapy. This would include agents added in vivobefore, during and after the apheresis process. There is no one targetor single strategy that must be pursued. Instead, applicant identifies alist of potential strategies that might be pursued.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated herein and constitutepart of this specification, illustrate exemplary embodiments of theinvention, and, together with the general description given above andthe detailed description given below, serve to explain the features ofthe invention.

FIG. 1 is a broad schematic illustration of the device of thisinvention, reflecting separation of whole blood into blood and plasma,treatment of the plasma, return of the plasma to the blood component andrestoration of the treated whole blood to the patient.

FIG. 2 is an illustration schematic of the portion of the device whereplasma is separated out of the blood, (which can occur via one ofseveral existing separation technologies) treated in at least one columnand returned via a conduit to the blood and donor.

FIG. 3 is a schematic of the inventive device deliberately echoing thestructure of FIG. 1, but reflecting the ability of this device tointroduce multiple columns and ports to both remove and add a variety ofcomponents to the plasma and ultimately the patient.

DETAILED DESCRIPTION OF THE INVENTION

This invention introduces a new method of addressing disabilities andconditions with interrelated strategies that improve the effectivenessof each strategy by reducing or eliminating other body systems and invivo responses which tend to limit the effectiveness of conventionalremedies. As only one example, a variety of medications and treatmentsare known to address excessive inflammation. Many are of limitedeffectiveness because of the mammalian tendency to induce inflammationfor a variety of reasons. One of the compounds mediating inflammation isGal-3. A conventional strategy to address inflammation may be made moreeffective by combining it with Gal-3 removal. Other treatments, whichmay be made more effective by reducing Gal-3 are disclosed. Gal-3 is anessential lectin expressed by the mammalian system, however, and cannotbe blocked entirely without morbidity and mortality. A system whichreduces Gal-3 concentrations while simultaneously administering othertreatments (either removal or addition of other agents) ordinarilycountered by effects mediated by Gal-3 in the body would be of value inthis one example. Other examples abound. Thus the invention is a methodfor treatment, and a system for treatment, which relies on apheresis,conducted through a system which withdraws a patient's blood, optionallyseparates it into cellular and plasma components, and treats both asrequired in separate, interchangeable modules the function of which isdictated by the individual patient's needs. A new form of “personalizedmedicine” which makes use of a number of interchangeable modules adaptedfor a particular patient's needs is provided.

Summary of New Apheresis Strategies

-   -   Strategic removal/modulation of multiple classes of bioactive        compounds by using a variety of specific and/or non-specific        filters.    -   Therapeutic control of levels of factors related and unrelated        to removal of Gal-3.    -   Modulation of any bioactive compound that can contribute to the        pathogenesis of any disease with utilization of the appropriate        filters.    -   Treatment individualized for patient as well as each treatment        optimized in relation to the individual's entire treatment        program.    -   Use of apheresis as a stand-alone therapy or in combination or        sequence with other pharmaceuticals, natural compounds,        botanicals, nutrients, vaccines, photosensitizing agents, immune        cell therapies, oxygen or oxidative therapies, ultraviolet or        other electromagnetic therapies, heat therapies, chemotherapy,        radiation, and surgical procedures with the aim of enhancing        effects, synergizing treatments, reducing side effects and        required dosages, and limiting longer term sequellae.    -   Use of apheresis in the acute setting to rapidly modify damaging        internal pathophysiological processes as well as provide for        rapid administration of specific therapies.    -   Use of apheresis in a preventive manner for conditions such as        diabetes that are caused by overreaction of the body to the        presence of viral infection, etc.    -   Use of apheresis as a periodic treatment for patients at high        risk of cancer metastasis or recurrence by removing factors that        increase risk and/or promote and/or enhance cancer growth,        and/or enhance cancer resistance, example being growth factors        such as VEGF, EGFR, TGF-Beta, etc.    -   Use of apheresis as a periodic treatment for a wide range of        chronic conditions, such as autoimmune conditions for which        control rather than cure is the goal, to prevent exacerbations        (which can be life-threatening in some cases), and limit        progression such as in chronic kidney disease, rheumatoid        arthritis, etc.    -   Use of this new apheresis capability to develop/and use in        conjunction with existing immune therapies (ability to collect        various immune cell components, modulate in various ways by        adding compounds, removal of compounds, multiplying the cells,        etc.), (such as CAR-T genetically engineered cell therapy).    -   Development of Circulating Tumor Cell (CTC) therapies by        collecting CTC's for both diagnostic and therapeutic purposes.    -   Development of and in conjunction with stem cell therapies,        dendritic cell therapies, synthetic antibody mimics, immune        checkpoint therapies, etc.

Traditionally, apheresis has been used to selectively remove a very fewclasses of agents or compounds. In the prior art, in general, molecularfilters were provided to exclude a whole class of molecules that cannotpass through a specific molecular sieve—generally referred to asmolecular weight range exclusion. U.S. Pat. No. 3,625,212, Rosenberg andU.S. Patent Publication No. 2006/0129082, Rozga are examples of thistechnique. It is not necessary, a priori, to exclude an entire class ormolecular weight range to make effective use of apheresis. Antigens,antibodies, and a variety of targeted binding agents, generally exposedto the passage of blood or plasma within the apheresis device, asdisclosed in pending PCT Application No. PCT/US14/38694 filed May 20,2014, can be employed to selectively remove one or more specific typesor classes of target compounds. Thus, this invention embraces a widevariety of apheresis removal strategies. Throughout this application theterm mammal is used to address particularly humans, but may also embracecommercial mammals (pigs, cows, horses, etc.) as well as companionanimals (dogs and cats, primarily). A few aspects of these are set forthbelow.

Compounds for Removal Using the Apheresis Process and Device for thisInvention Include

-   -   Cytokines, especially inflammatory cytokines    -   Cytokine receptors    -   Gal-s    -   Other Lectins    -   Growth factors    -   Growth factor receptors    -   Other growth factors    -   Inflammatory factor receptors, an example being TNF Alpha        receptors.    -   Inflammatory, compounds, precursors, metabolites, etc.    -   Tumor markers    -   Endopeptidase Metzincin superfamily    -   Tyrosine kinases    -   Other proteins involved in cell migration and metastasis    -   Other proteins involved in inflammation, fibrosis, and        degenerative diseases including    -   Advanced glycation end products,    -   Beta amyloid, alpha synuclein and others involved in        neurodegenerative diseases    -   Other proteins that can bind to essential nutrients, or can        block essential physiological processes and endocrine processes.    -   Immune complexes    -   Antibodies, examples include anti-DNA antibodies,        antiganglioside antibodies and other antibodies to both internal        tissues and pathogenic organisms.    -   Immunoglobulins of the different classes—IgG, IgM, IgA, IgE    -   Antigens    -   Cryoglobulin    -   Rheumatoid factor    -   Angiogenic factors such as polypeptides    -   Others including mycotoxins, viruses, bacteria, parasites and        their toxins.    -   Mast cells    -   Amyloid beta(40) and amyloid beta(42)    -   Antibodies to beta cells    -   Circulating tumor cells    -   Heavy metals and toxins a wide array of chemicals including        pesticides, dioxin, PCB's, BPA and other plasticizers, etc.—both        water soluble and fat soluble toxins.    -   Pro-inflammatory factors    -   Hormones and hormonal modulators    -   Neurotransmitters and neurotransmitters modulators

As noted above, the invention embraces not only removal of specifictargets or compounds, but addition of specific compounds to the plasmaor blood cells before return of the same to the patient's body. Often anaddition in this fashion allows drugs or other agents to be addedwithout further issues of patient compliance, without addressingquestions of interference, reduced absorption, bioavailability orlimited tolerance due to digestive tract kinetics or side effects, orbinding compounds in the patient's circulation, and the like. Inaddition, by adding agents to the blood stream while conductingapheresis to remove agents, a highly personalized, sequential andoptimized program of treatment can be effected through one step at onetime, rather than following a standard protocol of one treatment regimenfor every patient, or requiring patients to travel to multiple centersto receive multiple treatments. Often, agent addition can be balanced orselected to complement other agent addition, or blood borne compoundremoval through apheresis. A few of the agents that may be effectivelyremoved through the use of this invention are described above. A fewthat may be selectively added are listed below. These are typicallybioactive compounds that are added to achieve specific treatment goalseither as stand-alone therapy or in conjunction/sequencing with othertreatments (such as chemotherapy) and/or integrative therapies asdescribed above.

Addition of Bioactive Compounds

-   -   Recombinant Cytokines (that are being used as drugs)    -   Cytokines    -   Angiostatic proteins    -   Oxygen therapies    -   Stem cell therapies    -   Immune cell therapies    -   Hematopoietic cells of all kinds—B, T, etc.    -   Attenuated and/or modified and/or enhance cells, such as T        cells, B cells, macrophage, NK Cells, etc.    -   Dendritic cell therapies    -   Anti-fungal/anti-bacterial/anti-viral cytotoxic therapies    -   Heat shock proteins    -   Survivin    -   Immunoglobulin    -   Anti-CD20 antibody    -   Lymphocyte depleting antibody    -   Other immune stimulating factors    -   Agents for disrupting biofilm    -   Growth factors of various categories.    -   Anti-oxidants    -   Mineral, vitamins, nutrients, botanicals, proteins, amino acids.    -   Detoxification support agents, examples being Glutathione,        Taurine    -   Chelating compounds such as EDTA, DMPS, DMSA, Tetrathiomolybdate        (TM), etc    -   Pharmaceutical agents of different kinds, example being        chemotherapeutic agents, biological agents, hormonal agents,        etc.

The invention is more than a list of compounds that may be effectivelyselected for addition or targeted for removal using the apheresis deviceof the claimed invention. It is a new approach to apheresis, includingmodulation of either the plasma and/or solid components of blood, ortreatment of whole blood without separation, (without addition of eitherdonor or artificial plasma) that is based on the idea that eachindividual can benefit from a treatment model and protocol that istailored for the individual. In some cases, it might amount to no morethan some diagnostic testing to determine appropriate levels of thetarget compounds, which permit accurate determination of the number andtype of cartridges or filters the plasma should pass through, or theconcentration of the additives to be introduced prior to retransfusion.In other cases, however, where the targets and/or additives arespecific, or require a particular combination for a particularindividual, it may be advantageous to design specific filter cartridgesfor that individual. One example is the selective removal of cellsreflecting a genetic mutation in an individual, or harvesting stem cellsbearing a particular signature for modification for later return to theindividual to achieve tissue growth or removal or separation of cellsfor reintroduction with various immune therapy applications, etc.

In order to achieve the type of particularized treatment protocolscalled for by this invention, it may be necessary to formulateindividualized columns or cartridges which may then be switched in andout of the general apheresis device, as described in the referencedpatent application PCT/US14/38694 filed May 20, 2014. Indeed, while sometherapies may be acute or one time only, for those patient's with apersistent condition or disease state—cancer, auto-immune diseases andthe like, this invention allows preparation of an “ordered” set offilters to have on hand at facility proximal to the patient. Thisreduces costs and delays, and removes some of the anxiety associatedwith treatment. This opens up a variety of new designs and strategiesfor apheresis filter or column development not previously practicable.For instance, in prior art devices, a “one size fits all” approachgenerally required a molecular sieve strategy. In the new invention,even if patient one, for example, requires removal of Gal-3 and anauto-immune associated antibody, and patient 2 requires removal of Gal-3and an inflammation-associated cytokine, they both require the Gal-3removal filter, and wind up reducing the per patient cost ofdesign/development/manufacture of that common module then each has aspecific filter or column prepared for removal of the component, whichis specific to them. By sharing costs where possible, and using new lowcost technologies to make individualized patient care practical, newtreatment modalities are possible. A variety of strategies for columnconstruction are identified below, which make use of new technologies,such as 3D printing of columns and capture agents, such as antibodies,peptides, antibody fragments, aptamers, chemicals, antigens orderivatives, as well as the ability to tailor treatment of a particularindividual. Some of these are identified below:

Column Development Strategies

-   -   Pre-made columns based on a variety of technologies    -   Custom columns using basic materials that are FDA approved for        safety    -   Use of 3 D Printing technology particularly for custom columns        with patient specific matrices as well as for other more        generalized column structures

Column Use for Apheresis Procedure

-   -   Choice of column (s), number of columns, order of sequence    -   Timing Pretreatment, during other treatments, post treatments

Columns Designed for Processing of Various Blood Components

-   -   Use of whole blood    -   Use of plasma    -   Use of separated solid blood components (cells, larger proteins,        etc.)    -   Use of separators for specific cell populations

Clearly then, the new approach to preventative and therapeutic treatmentmade possible by this invention's system may be targeted to theindividual and adapted to the individual's needs, but relies in part onapparatus and practices and strategies applicable to entire groups andclasses of patients. A “column” is a traditional term to refer to asection of the device that may be switched in and out. Examples includea passage lined with antibodies that bind Gal-3 or a passage lined withantibodies that bind TNF Alpha, etc. Applicant recognizes that theactual design of the passageways, as well as the agents that bind targetcompounds, will change as personalized treatment comes to the fore (forexample, targets that are more difficult to bind might require a moretortuous path to extend the time they are in contact with bindingagents). New applicable materials and methods for column constructionwill become available. In a preferred embodiment, multiple different“columns” are designed and switched out as the patient responds totreatments. Thus, the filters or removal columns and modules becomeinterchangeable,

and the pattern and number of such “columns” can be varied from patientto patient or for the same patient at different times using the samemachine. For example, in a patient needing removal of only one targetcompound, several “column” spacings might be occupied by straightunlined tubing, where the same machine when used for another patientmight exhibit 3 or 4 or more “columns” placed in sequence. Ultimately,the system becomes one where there is a basic machine, with spaces forthree or four or five “columns” or filter packs. Each module site may beoccupied, or there may be capability for bypass using appropriate tubingand connectors when not occupied by an active column. In suchcircumstances, not only column design and character, but sequencing ispart of the individualization of treatment. Different issues to takeinto consideration in developing patient strategies are set forth below:

Basic Concepts

Columns

-   -   Pre-made columns: Columns remove specific components via a        variety of potential technologies including selective removal,        size exclusion, non-selective removal, receptor binding,        antibody binding, precipitation, charge based, size exclusion,        electromagnetic, etc.    -   Custom columns: Obtain approval for the basic materials/FDA        approved safe materials, and/or materials that are inert and        will not cause an adverse reaction in the binding process.        -   Custom features made to order based on individualized            assessment, which could include analysis of: tumor            genetics/phenotype and membrane receptors, levels of            elevated blood borne cancer and inflammation promoting            factors (CIPFs), immunological factors, etc. described more            fully below.        -   3D printing technology and other technologies could include            patient specific tissue matrices with embedded components            both biological and electrical such as sensors to detect            concentrations, so that the flow through the columns and            concentrations of agents, cellular components and CIPFs            could be monitored and modulated appropriately. Custom            columns/filters developed that would be suitable for            treating any biological condition including inflammatory,            degenerative, allergic, autoimmune, infectious, fibrotic and            neoplastic.

Sequencing

Of Columns

-   -   choices of columns    -   numbers of columns    -   order of sequence

Of Timing

-   -   Apheresis pre-treatment (such as prior to chemotherapy) or        during treatment (for example during chemotherapy)    -   Post treatment

Plasma Separation as Well as Whole Blood Filtering Systems

-   -   With use of whole blood, once separated, the two components can        be worked with simultaneously and then reunited, or different        procedures can be accomplished using one or the other (i.e.,        meaning plasma, whole blood, or blood cellular components)        depending on the goal of treatment.    -   With use of unseparated whole blood, specific cell types can be        removed or treated as well as modulation of blood borne proteins        and other factors. Whole blood that has been cleaned using        plasma apheresis has lipids and inflammatory compounds removed        which would enhance the detection of CTC's, immune cells, etc.,        and would thus be a preliminary treatment to further apheresis        applications as described in this document.

Development of Columns Based on 3D Technology

This new technology is just beginning to reveal its capabilities forconstructing scaffolding, matrices, living tissues, etc. that can beenvisioned as material for column construction for specific purposes.These columns would be designed for a variety of complex functionalcapabilities with examples further described below.

-   -   Optimized for removal of one or more specific factors either in        combination with Gal-3 removal or independent, with option to        use multiple filters in a cascade to remove multiple compounds.    -   3D printing of receptors, matrix embedded compounds with high        affinity for specific factors. For example, the printer could        print a column that contains receptors for growth factors,        signaling factors, etc., including but not limited to VEGF,        EGFR, TGF beta family, interleukins, fibroblast growth factors,        neurotrophic factors, cancer cell receptors (CD family)        receptors for various galectins, survivin, etc.    -   3D Printing of specific binding sites for individualized columns        based on:        -   Laboratory testing        -   Profiles of tumor expression        -   Genomic analysis of the tumor,        -   Genomic analysis of the individual.        -   Analysis of the tumor cell membrane characteristics and            protein expression    -   Materials used will be FDA approved as safe, etc, or inert        compounds that do not interact.    -   Filter construction, either using 3D printing or other design        may bind or attract based on a number of different technologies        including physiological binding antigen/antibody, receptor        binding (for example EGFR), electric charge, size exclusion,        etc.    -   Incorporation of synthetic antibody mimics (SyAMs) and fragments        (e.g., scFv, Fab, Fab′, Fc) which attach themselves        simultaneously to disease cells and disease-fighting cells.        Included within this approach is the addition of various peptide        fragments such as Fab, FC, etc.    -   Potential for incorporation of electrical sensor technology        which could be used for identifying particular molecules or        sensing levels of molecules.    -   Columns might include UV, laser, or other light capability,        (existing photopheresis technology), or magnetic fields.    -   UV light, laser, light sources and energy attenuation devices        (EMF's and other energy sources) can be integrated in to the        extracorporeal apheresis procedure, in all stages: Whole blood,        separated cells in general and subpopulations, plasma, joined        blood (cells and plasma) both separation. The goal here is to        make it specific to source of energy and frequency, and to make        it timing specific as well.    -   Electromagnetic therapy could be in the form of a single column        or embedded in the matrix of a column, such as one that could        also bind CTC's, bacteria, viruses, spirochetes.    -   For example: a matrix that would attract and bind specific        organisms, along with electronic sensing imbedding to monitor        density. UV or other therapy could then be turned on based on        this to destroy accumulated organisms.    -   This could be combined with a column that could be made to bind        specific compounds that act as growth promoting substrates for        infectious organisms.    -   Photosensitizing medications can be incorporated.    -   Cancer cells could also be targeted with this type of        technology. 3D Matrix could contain binders for CTC as a way to        remove, or capture these cells to introduce T-cells or other        immune cells reactive to the tumor for activation, right in the        column instead of removing from the body altogether and        reintroducing as is done currently.    -   Alternatively, personalized materials may be harvested and        cultured, but reintroduced in the apheresis process to ensure        reduced side effects and increased efficacy. For example, by        reducing pro inflammatory cytokines and compounds and adding        other compounds that will enhance the therapeutic effect or        improve the delivery to the target tissue, apheresis can be used        to improve the effectiveness of other therapies.    -   Apheresis can be used as a purely diagnostic exercise (As one        example, CTC's might be bound and held on a 3D scaffolding that        can be separated from the patient and used to test chemo drugs        and other therapies for efficacy).    -   Compounds can be run through a column where particular cell        components or other factors have been bound, with the purpose to        “unglue bound components at a strategic point for the next step        in the therapy.    -   The process could be continuous, or could be an adjunctive        procedure which could be another part of the machine that would        siphon a certain quantity of whole blood, and cycling it thru a        column over some hours with heparin where the cells are cultured        in the column and returned, so you can have a process where you        are culturing a treatment on one side and you are removing the        inflammatory compounds at the same time    -   Using 3D technologies to construct custom scaffolding on which        living cells can be cultured producing tissue matrices such as        connective tissue (using NAG or other compounds), which can        attract binding by Lyme spirochetes or other infectious agents,        making the spirochetes accessible for therapies including T-cell        stimulation therapies, allowing for removal or for direct        application of light or other cytotoxic therapies. Tissues could        be made homologous to the patient and printed on columns for        various uses such as above    -   Detection of subpopulations of cells for example RBC's that are        infected with Plasmodium falciparum (Malaria), Babesia spp.        (Babesiosis), and alike, allowing for elective removal of the        infected cells without causing anemia and too big of depletion        of red blood cells. Subpopulations of cells can be targeted.    -   Cells can be grown into a type of matrix or tissue that can have        other technologies imbedded in it such as combination of the        tissue with electronics, sensors, etc.    -   Electronic sensing capabilities can be used for a multitude of        purposes in terms of sensing levels of specific compounds        passing through the filter, or the concentration of bound        compounds, or density of particular cell types bound to filters,        or microorganisms. This can allow for timing of the various        components of the particular procedure.

Immune Therapies and Circulating Tumor Cell Therapies

To develop new technology for advanced immune therapy based on filtersand devices such as centrifugation procedures with the capability tocollect various relevant cell types including circulating tumor cells(CTCs), T-cells, B-cells, macrophages, monocytes, dendritic cells, andother emergent immune modulating concepts are listed below.

-   -   CTC collection for therapeutic and diagnostic purposes.    -   Cells could be analyzed ex-vivo for genetic and membrane        specific receptors, proteins, as well as their secretion of        cytokines and other factors such as IL-6 and IL-8.        -   For example, if we find that a person's CTC's have a            particular receptor, e.g., CD4, we can target them with            binders for this receptor.    -   CD receptor family, CCR receptor family.    -   We can create a custom column containing one or multiple binders        for patient specific CTC characteristics.    -   Can harvest CTC's and hold them on a 3D scaffolding that can be        separated from the patient and used to test chemo drugs and        other therapies for efficacy. (Organovo).    -   Patient specific therapies based on the results would be        developed.

Incorporation of Targeted Stem Cell Therapies

Binding of CTC's and or T/B cells for purposes of “educating” the immunesystem is now possible. Identifying Cancer stem cells (CSCs) andremoving them specifically, or creating therapies targeting thereceptors in the CSC's promoting therapies each made more effective bytheir combination. These are among the principal reasons for recurrencesand resistance to therapy.

-   -   The bound agents and materials can be reinfused at a later date.        Alternatively they could be co-cultured on a column with whole        blood being circulated. This would take a certain amount of        time. Example: you have an antigen specific to the cancer cells,        and you have B cells co-cultured, then you run heparin or an        appropriate agent that will release the cells at a certain        point. The B cell attaches to the receptor and the antibody or        FC fragment gets stimulated, then the potentiated B cells are        re-infused in a closed system.    -   Using apheresis to change the density of the antibodies to        control the rate of exposure and the rate of the reaction        Examples of Targeted Stem Cell Therapies that could be        Incorporated into this Technology:    -   CAR-T: T-cells genetically engineered to produce special        receptors on their surface called chimeric antigen receptors        (CARs). CARs, proteins that allow the T cells to recognize a        specific protein (antigen) on tumor cells.    -   Engineering stem cells to secrete cancer-killing cytotoxins.        These stem cells are placed inside the tumor and destroy the        tumor from within. The cytotoxins can be targeted to cells with        specific receptors such as epidermal growth factor or IL-13        receptor alpha 2 (IL13RA2) found in many brain tumors (this work        is in process).    -   Dendritic cell therapy    -   Targeting activation and regulation of T cells    -   Oncolytic virus vaccines    -   Adoptive T cell therapies    -   Collection, identification, and selective removal of the        subpopulation of immune cells that are excreting specific        cytokines (for example inflammatory IL-6 and IL-8).    -   Incorporation of synthetic antibody mimics (SyAMs) either as a        therapy in itself or incorporated into the filter technology.        -   SyAMs work first by recognizing cancer cells and binding            with a specific protein on their surface. Next, they also            bind with a receptor on an immune cell. This induces a            targeted response that leads to the destruction of the            cancer cell.

This invention not only opens up new opportunities to personalizetreatments for a specific mammalian patient but allows the complementingof other treatments the patient may be receiving. Thus, often certaintreatments will aggravate or unnecessarily suppress natural bodyresponses, like inflammation or stomach or bowel upset, due to a cascadeof conditions or comorbidities that are mediated by factors other thanthe one directly involved in the patient's condition or therapy. Otherphenomena that may be associated with treatments involving surgery andthe like may include the development or aggravation of fibroses, orupregulation of growth factors, increasing risk of metastasis, etc. Suchcombined therapy approaches will require consultation and comprehensivemonitoring, but may vastly increase the effectiveness of traditionaltreatments. Accordingly, part of this invention is employing apheresisto remove agents or conditions that that might otherwise interfere withother treatments. Some of these strategies include a variety of uses ofapheresis and selective targeted removal associated with othertreatments including the following concepts and approaches:

-   -   Use Apheresis Technology to therapeutically control levels of        circulating CIPFs and immune modulating factors, all of which        play a role in different conditions and could be removed or        modulated at different stages, both related and unrelated to        removal of Gal-3 based on the patient and the condition being        treated. Apheresis becomes like a control device where you make        the adjustments outside of the body instead of inside the body        where the inflammatory response can be much more difficult to        control.    -   Regulate the inflammatory response to reduce side effects of        standard therapies as well as emerging targeted treatments,        enhance effectiveness, reduce side-effect induced treatment        interruptions, and allow for lower, more tolerable doses of        agents used.    -   In preparation for, prior to, in conjunction with, and/or        post-surgery, chemotherapy, pharmaceutical therapies, targeted        therapies such as mAB, immune therapies, hormonal therapies,        radiation, hyperthermia, photodynamic therapy, and other        integrative therapies using botanicals, nutrients, etc.    -   Reduce pro-metastatic inflammation and the upregulation of        growth/repair factors potential for release and growth        stimulation of tumor cells post-surgery.    -   Removal of CTC's pre and/or post-surgery so you don't let the        body induce a metastatic process especially with tumors that        spread hematologically.    -   Reduce post treatment fibrosis secondary to surgery and        radiation.    -   As a result of chemical, pathological or chronic inflammatory        insult to an organ or organs such as is the case with chronic        kidney disease, liver diseases, lung diseases, cardiovascular        diseases, and exposures to toxins such as pharmacological        agents, mycotoxins, heavy metals, etc.    -   Reducing inflammatory compounds that protect the tumor cell        environment from access by chemotherapeutic and immune        stimulating agents.    -   Addition of other agents using sophisticated protocols in        combination with columns, such as IV vitamin C, artesunate,        ozone, honokiol and its derivatives, oxidative therapies,        enzymes, heparin, other anti-viscosity agents, photosensitizing        agents, pharmaceuticals, chelating agents, ultraviolet and other        light and electromagnetic therapies, etc.    -   Reducing growth factors in the blood to enhance the        effectiveness of growth factor inhibition therapies such as        tyrosine kinase inhibitors, etc.    -   Agents specific for disrupting biofilm, the protective sheath        that covers tumors, arteriosclerotic plaques, Lyme spirochetes,        autoimmune processes, etc.

The invention further comprises introducing agents to provide a noveltreatment for infectious diseases such as Lyme disease, for example, byusing compounds that can cause an infectious organism, such as Lymespirochetes, to move from tissue sequester sites into the bloodstream.This could be combined with aforementioned filter to simultaneouslyremove growth factors specific for the infectious organism as well ascolumn matrices (which could be living tissues) that would attract saidspirochetes for binding, followed by in column cytotoxic therapy,essentially sequestering the cytotoxic treatment from the patient'ssystem, reducing systemic reactions and side effects.

Among the most intractable and devastating of diseases are a host ofauto-immune diseases. The invention specifically contemplates usingfilter technology to create novel sequential strategies for addressingautoimmune diseases in a similar fashion. Many autoimmune processesoccur in pockets, the body walls off sites of infection, the organismscreate protective biofilm, creating an anaerobic and/or isolatedenvironment that the immune system cannot access. This creates a chronicinflammatory locus which can develop into an autoimmune process whichwill stop when the area is exposed and treated.

Introduction of oxidizing agents once infectious sites have beenaccessed using anti-biofilm therapies. Example: manipulatingmyeloperoxidase, which is bound by heparin (and spikes during LDLapheresis) time release to enhance a specific antimicrobial therapy. Maychoose not to remove elevated Gal-3 and/or treat with inflammatory oroxidative compounds to enhance effect for infectious diseases whilecontrolling other inflammatory cytokines.

Gal-3 could be retained during certain points of therapy for fightinginfectious organisms due to its enhancing effect. It might be removed atthe end of treatment to reduce inflammation produced by the process ofantimicrobial therapies. Other galectins may be retained or removed atany stage as only a few examples, galectins 1 and 9 have been implicatedin rejection syndrome, and in impregnation difficulties.

Introducing agents, such as growth factor inhibitors, or multi-drugresistance (MDR) inhibitors along with standardized therapy or justprior to or following to enhance effectiveness of anticancer therapiessuch as chemotherapy. These agents could be introduced at key pointsduring or following apheresis.

-   -   Remove specific growth factors while retaining other's needed to        balance the reaction.    -   Increase the effectiveness of immune therapies by reducing        inflammation, treatment side effects and improve the response by        allowing the cells to get to the target tissue much easier.    -   Incorporating new classes of immune therapies such as targeted        oncovirus, T and B cell therapies, dendritic cell therapies,        stem cells, nanotechnologies for targeting tumor cells and tumor        macrophages, etc., immune modulation agents, such as PD-1/PD-L1        inhibitors (antibodies that allow the immune system to attack        tumor by blocking the pathway that paralyzes the immune        response).    -   To use instead of/or reduce the dosages of existing        pharmaceutical treatment for diseases such as autoimmune        conditions by binding to inhibitory proteins, antigens and        circulating immune complexes that drive tissue damage. Therapy        could reduce or eliminate the need for immune-suppressive        therapies with their negative long term side effects.    -   As a periodic treatment for patients at high risk for metastasis        or recurrence to remove both inflammatory and hyperviscosity        factors and CTC's that may be putting them at risk.

The use of the apheresis invention disclosed herein has been describedabove in the context of individualized treatment of mammalian and humanpatients, and as an element of a combination of therapies for suchpatients. Thus, apheresis may be effective on its own in addressing orreversing a variety of disease states. This invention embraces a varietyof strategies to personalize such treatment, to tailor it for the needsand character of the individual patient. It may also be effectively usedto promote the effectiveness of other therapies, such as cancertreatment and the like. It may also be used, as discussed above, toremove factors that would otherwise inhibit the effectiveness of moreconventional strategies, or make it possible to enable administration ofthose strategies without profound risk, such as the risk of fibroses ortumor metastases. It is not limited, however, to therapeutic or clinicalintervention. It finds application in pre-clinical or pre-therapeuticsituations, or when the patient is not yet symptomatic. Some of thesepre-clinical applications include:

-   -   Removal of inflammatory compounds, immune complexes and        antibodies in advance of the overt clinical manifestation of        autoimmune diseases.    -   Removal of inflammatory and autoimmune complexes in other        preclinical situations, and specifically in IDDM, or Type I        diabetes. Given prescreening in high risk for Type I Diabetes        and screening for antibodies against langerhans cells (Beta        islet cells of the pancreas) apheresis may be used        pre-clinically to prevent development of diabetes. Gal-3 is        implicated in the inflammatory process in diabetes. Gal-3        removal coupled with removal of other complicating factors may        suppress the disease or its most debilitating symptoms.    -   Remove the antibodies via apheresis. There is elevation in these        antibodies long before type I diabetes sets in.    -   Couple this with the various anti-inflammatory therapies        discussed above.    -   Remove the viruses that are implicated in the initiation via        apheresis.    -   If the typical immune inflammatory response is avoided using        apheresis, diabetes can be prevented.    -   This strategy will also apply to other autoimmune conditions,        myopathies, neurodegenerative diseases, etc. such as ALS, MS,        Alzheimer's, Parkinson's and other degenerative diseases that        are devastating in their outcome and extremely expensive to        treat once established.    -   While the use of therapeutic apheresis is already known for some        of these diseases, such as Crohn's disease, neuromuscular        conditions, etc. to the best of our knowledge, it has not been        used in a pre-clinical or pre-symptomatic situation, to prevent        development of this disease condition.

Fundamentally, this invention opens new opportunities and avenues forthe use of apheresis, which may include blood cell treatment, plasmatreatment or both. It permits the personalization of treatment withoutextraordinary expense by deploying a single device or apparatus that canbe modified for individual treatment by the switching in and out oftargeted columns or filters without the cost of development of a newmachine for each major type of illness or condition. It allows for thesimultaneous maintenance of conventional technologies that may otherwisebe limited or prohibited because of responses due to removable targets.It makes apheresis of value in both clinical and pre-clinical settings.However apheresis is only as good as the effectiveness of the column(s)in removing the target compound or subject matter. The development ofunique antibodies and binding partners for a universe of materials, andthe ability to deliver or “print” these on a scaffold or supportingsurface over which a patient's blood or plasma can be led, vastlyexpands the capabilities and applications that apheresis may be put to.We identify below only a handful of the types and categories of targetcompounds and subject matter that may be removed by the “personalized”apheresis of this invention. At least ten different very broadcategories, as well as a “miscellaneous” category may be identified, asorganized below: Many of these compounds are multifunctional and havecomplex physiological roles, with necessary regulatory functions inspecific conditions and situations, while in other situations would havemore pathogenic effects warranting removal.

Cytokines

A large group of soluble extracellular proteins or glycoproteins, keyintercellular regulators and mobilizers.

-   -   Classes of Cytokines:        -   Chemokines        -   Homeostatic: are constitutively produced in certain tissues            and are responsible for basal leukocyte migration. These            include: CCL14, CCL19, CCL20, CCL21, CCL25, CCL27, CXCL12            and CXCL13. This classification is not strict; for example,            CCL20 can act also as pro-inflammatory chemokine.        -   Inflammatory: these are formed under pathological conditions            (pro-inflammatory stimuli, such as IL-1, TNF-alpha, LPS, or            viruses) and actively participate in the inflammatory            response attracting immune cells to the site of            inflammation. Examples are: CXCL-8, CCL2, CCL3, CCL4, CCL5,            CCL11, CXCL10, CCL's, CXL's, CX3CL, XCL'S        -   Interleukins-multiple subclasses (IL) and families some of            which are proinflammatory        -   Interferons (IFN)        -   Lymphokines        -   Tumor necrosis factor families (including TNF-alpha, -beta)        -   Adipokines (adipocytokines): Cytokines secreted by adipose            tissue.        -   Leptin        -   Adiponectin        -   Apelin        -   Chemerin        -   Interleukin-6 (IL-6)        -   Monocyte chemotactic protein-1 (MCP-1)        -   Plasminogen activator inhibitor-1 (PAI-1)        -   Retinol binding protein 4 (RBP4)        -   Tumor necrosis factor-alpha (TNF-alpha)        -   Visfatin

Other Cytokine Related Targets

-   -   Receptors: removal increases response at target tissue.        -   Chemokine Receptors        -   CCL13 (MCP4), CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR8,            CX3CR1, CXCR1 (IL8RA), CXCR2 (IL8RB).        -   Interleukin Receptors        -   IL10RA, IL10RB, IL1R1, IL5RA (CD125), IL9R.        -   Other Cytokine Receptors        -   TNFRSF11B (OPG).        -   Galectins        -   Lectins        -   Growth Factors:            -   Epidermal Growth Factor Receptor (EGFR)            -   Epidermal Growth Factor (a kinase)            -   Activator Protein-1 (AP-1)            -   Vascular endothelial growth factor (VEGF) family            -   Hepatocyte growth factor (HGF)            -   Transforming growth factor alpha family and beta family            -   Basic fibroblast growth factor (bFGF) (-2)            -   Tumor necrosis factor alpha (TNF alpha)            -   Heparin binding epidermal growth factor            -   Insulin-like growth factor (IGF) family including IGF 1,                2 Vascular adhesion molecule-1            -   Endothelial intercellular adhesion molecule        -   Other Growth Factors Include:            -   Adrenomedullin (AM)            -   Angiopoietin (Ang)            -   Autocrine motility factor            -   Bone morphogenetic proteins (BMPs)            -   Brain-derived neurotrophic factor (BDNF)            -   Erythropoietin (EPO)            -   Glial cell line-derived neurotrophic factor (GDNF)            -   Granulocyte colony-stimulating factor (G-CSF)            -   Granulocyte macrophage colony-stimulating factor                (GM-CSF)            -   Growth differentiation factor-9 (GDF9)            -   Erythropoietin (EPO)            -   Healing factor            -   Nagalase            -   Hepatoma-derived growth factor (HDGF)            -   Sonic Hedgehog (polypeptide-abnormal activation of the                pathway probably leads to development of disease through                transformation of adult stem cells into cancer stem                cells that give rise to the tumor)            -   Interleukins: 1-7            -   Keratinocyte growth factor (KGF)            -   Migration-stimulating factor (MSF)            -   Myostatin (GDF-8)            -   Nerve growth factor (NGF) and other neurotrophins            -   Thrombopoietin (TPO)            -   Wnt Signaling Pathway            -   placental growth factor (PGF)            -   Foetal Bovine Somatotrophin (FBS)            -   Renalase-RNLS-Anti-apoptotic survival factor            -   Endothelins            -   Ephrins        -   Tumor Markers: (biologically active)            -   CEA (carcinoembyronic antigen)            -   CA-125 (cancer antigen-125)            -   CA-19-9 (cancer antigen)            -   CA 15-3            -   CA 27-29            -   Prostatic Acid Phosphatase            -   Prostate Specific Antigen (PSA), proPSA (p2PSA), freePSA        -   Endopeptidase Metzincin Superfamily.            -   Matrix Metaloproteinases: most important class MMP                Groups are the collagenases, the gelatinases, the                stromelysins, and the membrane-type MMPs (MT-MMPs).            -   Adamalysins, serralysins            -   Astacins            -   Collectively, these enzymes are capable of degrading all                kinds of extracellular matrix proteins, but also can                process a number of bioactive molecules. They are known                to be involved in the cleavage of cell surface                receptors, the release of apoptotic ligands (such as the                FAS ligand), and chemokine/cytokine inactivation. ^([2])                MMPs are also thought to play a major role on cell                behaviors such as cell proliferation, migration                (adhesion/dispersion), differentiation, angiogenesis,                apoptosis, and host defense. There are a wide variety of                MMPs, some are noted below.        -   Tyrosine Kinases:            -   soluble fms-like tyrosine kinase-1 (sFlt-1), (elevated                in preeclampsia)            -   sVAP-1 (serum vascular adhesion protein-1)            -   ICAM-1            -   VACM-1            -   Others        -   Proteins involved in cell migration/metastasis: (some of            these are newly discovered and are primarily intracellular,            but may be shown in future studies to also be found in the            circulatory system).        -   DENND2B: DENND2B activates another protein in the cell            called Rab13, which is an enzyme that promotes cell            migration,        -   RSK2: significantly increases cell migration in part by            reducing integrin activation. Integrins play an important            role in cell adhesion to their surrounding tissue and the            migration of tumor cells to new locations in the body. RSK            is active in both breast and prostate tumors, and promotes            proliferation in these cells. It can also promote cell            invasion and metastasis in head and neck cancers in addition            to lung cancer and neuroblastoma.        -   hnRNPM: helps launch a cascade of events that enables breast            cancer cells to break away from the original tumor,            penetrate the blood stream, invade another part of the body            and form a new nodule of that tumor.        -   HIF-1        -   JMJD2C        -   HIF-1 switches on the JMJD2C gene, stimulating production of            the protein. HIF-1's presence also enables JMJD2C to bind to            DNA at other HIF-1 target genes, and then loosen those DNA            sections, enabling more HIF-1 to bind to the same sites and            activate the target genes.        -   NOS2: high levels of NOS2 are a predictor of poorer survival            in patients with ER-negative breast tumors and to suggest            that selective NOS2 inhibitors might be of benefit to these            individuals. Inducible nitric oxide synthase (NOS2) is            involved in wound healing, angiogenesis, and carcinogenesis.    -   Other Proteins and Proteases:        -   Bcl-2 family        -   Inhibitor of Apoptosis family (includes survivin)        -   Cathepsin Family            -   A, B, C, D, E, F, G, H, K, L1, L2, o, S, W, Z            -   Integrins            -   Antigens            -   Antibodies            -   Immune complexes            -   Caspase family (most act intracellularly, but some are                found in serum)            -   Myeloperoxidase        -   Angiogenic Polypeptides: (which may be listed elsewhere)            -   Activator protein 1 (AP-1)            -   Angiogenin (AG) and angiotropin (AT)            -   Angiopoietin (APN)            -   Basic fibroblast growth factor (bFGF)            -   Cyclooxygenase (COX) and lipoxygenase (LOX)            -   Granulocyte-colony stimulating factor (G-CSF)            -   Hepatocyte growth factor (HGF)            -   Insulin-like growth factors 1 and 2 (IGF-1 and -2)            -   Interleukin-8 (IL-8)            -   Nuclear factor kappa B (NF-kB)            -   Placental growth factor (PGF)            -   Platelet-derived endothelial cell growth factor                (PD-ECGF)            -   Platelet-derived growth factor (PDGF)            -   Pleiotrophin (PTN)            -   Proliferin Thrombospondin-1 (TSP-1)            -   Transforming growth factor alpha (TGFα)            -   Transforming growth factor beta (TGFβ)            -   C Reactive Protein (CRP)            -   Tumor necrosis factor alpha (TNFa) and TNFr3 (has both                pro and antiangiogenic roles) Vascular permeability                factor (VPF)        -   Others:            -   Malondyaldehyde (MDA)            -   2-thiobarbituric acid (TBA)            -   Secretins            -   Adenosine diphosphate (SDP), thrombin and other                molecules that participate in platelet-tumor cell                interaction.            -   Tumor-derived platelet agonists, such as adenosine                diphosphate (ADP) and thrombin, induce platelet                activation, followed by the formation of heterotypic                aggregates that protect tumor cells from immune attack                and physical damage. Tissue factor expressed by tumor                cells also leads to thrombin generation through the                activation of the coagulation cascade that ultimately                results in fibrin formation and platelet activation.                These processes, among others, promote the development                of venous thromboembolism in solid tumor and                hematological malignancies.            -   Fungus and/or their toxins-Mycotoxins            -   Viruses            -   Microbial agents—bacteria, viruses, fungus, helminths,                parasites and their toxins

Also as noted, apheresis as employed in this invention can be, but neednot be, a “stand alone” treatment, even if not associated with someother treatment modality. Given the proper apheresis device, such asthat set forth in PCT/US14/38694 filed May 20, 2014, there are importantopportunities to supplement the blood of a patient without ever treatingthe patient's body, by adding agents to the blood or plasma stream exvivo. The added agents can be cells or cell fragments or compoundsharvested from the patient's body, altered and returned, or syntheticdrugs or compounds or agents new to the patient, including a variety ofnatural agents, and everything in between. The point of addition isflexible, depending on the agent to be added, it may be added before,during or after the whole blood is otherwise treated in the system.Although it is clear that from the date of this writing onward, newpotential agents to be added via the “return loop” of the apheresisdevice of this invention (that is, after the blood or plasma or both haspassed through the columns or filters) some are specifically mentionedbelow:

-   -   Addition of Cytokines during apheresis for various treatment        protocols could include: (Some cytokines have been developed        into protein therapeutics using recombinant DNA technology.        Recombinant cytokines being used as drugs as of 2014 include):        -   Bone morphogenetic protein (BMP), used to treat bone-related            conditions        -   Erythropoietin (EPO), used to treat anemia        -   Granulocyte colony-stimulating factor (G-CSF), used to treat            neutropenia in cancer patients        -   Granulocyte macrophage colony-stimulating factor (GM-CSF),            used to treat neutropenia and fungal infections in cancer            patients        -   Interferon α, used to treat hepatitis C and multiple            sclerosis        -   Interferon beta, used to treat multiple sclerosis        -   Interleukin 2 (IL-2), used to treat cancer.        -   Interleukin 11 (IL-11), used to treat thrombocytopenia in            cancer patients.        -   Interferon γ, used to treat chronic granulomatous disease &            osteopetrosis.        -   Addition of angiostatic proteins        -   Addition of factors in conjunction with chemotherapy        -   Addition of oxygen therapies including in conjunction with            hyperbaric oxygen treatment        -   Addition of bacterial and viral cytotoxic therapies        -   Addition of anti microbials of all classes—anti viral, anti            bacterial, and helmetic, anti parasitic.        -   Addition of Survivin            -   Survivin is a strong T-cell-activating antigen, and                clinical trials have already been initialed to prove its                usefulness in the clinic.        -   Addition of Heat Shock Proteins (HSPs) as cancer vaccine            adjuvant            -   Given their role in antigen presentation, HSPs are                useful as immunologic adjuvants in boosting the response                to a vaccine. Furthermore, some researchers speculate                that HSPs may be involved in binding protein fragments                from dead malignant cells and presenting them to the                immune system. Therefore HSPs may be useful for                increasing the effectiveness of cancer vaccines.                Extracellular located or membrane-bound HSPs mediate                immunological functions. They can elicit an immune                response modulated either by the adaptive or innate                immune system. In cancer, most immune therapeutic                approaches based on extracellular HSPs exploit their                carrier function for immunogenic peptides.

As noted above, Applicant's invention allows an unprecedentedopportunity to combine therapies that reinforce each other. One of theprincipal achievements using this therapy is the selective removal ofGal-3. By employing plasmapheresis or apheresis to selectively removeGal-3 (removal can be effected by removal from whole blood or fromplasma following separation) by even a limited percentage (for instance,ten percent) a significant improvement in both acute and chronicinflammation can be achieved. But by removing the blood from the patientand providing a limited time opportunity to selectively impact othercomponents of the blood, a variety of therapies which are interrelatedin terms of action and target can be combined in time as well, improvingeffectiveness. In terms of the drawings provided, the blood in such anembodiment is withdrawn and pumped by pump 1006 from mammal 1002. Ifdesired, it is separated at filter 1012 into blood components and plasmacomponents, although the whole blood may be treated as one unit. Theblood passes through module 1008. In this example, the module comprisesan opening through which the blood or plasma is led which comprises aGal-3 binding molecule the blood or plasma is exposed to. The molecule,an antibody, antibody fragment, peptide or polysaccharide adapted toselectively bind Gal-3, binds to Gal-3 as the blood passes through,showing a measurably reduced amount of Gal-3 after it has passed throughthe system and is returned to the mammal. Reductions on the order of atleast 10%, more effectively 20, 25, 40, 50 or even 60% of circulatingGal-3 may be effected simply by providing more than one module 1008 withGal-3 binding moieties.

One therapy augmented by the invention disclosed herein is immune cellactivation and regulation. As noted, Gal-3 is a mediator ofinflammation. Increasing evidence demonstrates, however, that Gal-3 isalso an important regulator and modulator of solid tumor environments,and anti-tumor activity. Thus, Gal-3 has been shown to modulate T-cellresponses to both non-solid and solid tumors, including pancreatictumors, through more than one mode of action—including apoptosis, T-cellreceptor (TCR) cross-linking and TCR downregulation. Kouo et al, CancerImmunol. Res., 2015, 3(4): 412-23. Thus, Gal-3 has been found elevatedin many solid and non solid-tumor cancers, and appears to support cancercell survival. Ruvolo, Biochim. Biophys. Acta, 2016, 1863(3):427-37.Accordingly, while activated tumor cell treatment is a recognizedanti-cancer treatment, the body's natural mechanisms, including Gal-3work to offset any treatment gains that might be achieved by immune cellactivation.

One method of regulating, augmenting and enhancing immune cellactivation in an ex vivo environment is photopheresis. In extracorporealphotopheresis, white blood cells (hereinafter “WBC’) are separated outand collected. While only a small fraction of a patient's total WBC areextracted for treatment in any one passage, a patient can go throughseveral rounds of treatment, which is consistent with the timing ofapheresis, which permits treatment for a period of hours. Inphotopheresis, the WBC are mixed with an intercalating agent, such as8-MOP (methoxsalen) and irradiated with UV light. The WBC are thenreturned to the body. Using the invention disclosed, this is simply onemore module 2008 that the patient's blood or plasma is passed through.The resulting leukocytes are returned to the body. Although thesetreated cells are more subject to apoptosis than untreated leukocytes,since only a small fraction of the patient's cells are treated in anysitting, their susceptibility to apoptosis alone does not account forthe effectiveness of the treatment. It also appears that the treatedapoptotic-prone cells are taken up by dendritic cells and macrophages.It is through interaction with the dendritic cell system that thetreated cells influence rebalancing of the immune system.

As noted, the body's systems, modulated at least in large measure byGal-3, tend to limit the impact of photopheresis. Thus, much of thebenefit of photopheresis is limited by the tendency of Gal-3 to protectthe tumor microenvironment the activated immune cells are to address.Gal-3 also suppresses the immune response and release of cytokines byimmune cells. Gal-3 is also frequently a pro-inflammatory driver, whichagain tends to limit the ability of in vivo blood treatments to impactsystemic or localized disorders in the body. Photopheresis is also usedin the treatment of a variety of diseases other than T-cell leukemia,including cutaneous T-cell lymphoma and in graft versus host disease.While these treatments are helpful, they are of limited impact becausethe body's relatively weak or incompetent immune system, oftenaggravated by inflammation, is unable to capitalize on the advantagesprovided, because its own system regulates to the contrary.

One example of the tendency of body systems mediated in part by Gal-3expression is in the role of the development of monocyte-deriveddendritic cells, the very cells whose expression is a target of theactivated immune cells discussed above. Gal-3 has been shown to beimplicated in the induction of a T helper 2 response, but little wasknown about its interactions. Gal-3 has been shown to inducephosphatidylserine exposure and apoptosis in dendritic cells and theirdifferentiation. Van Stijn et al, Mol. Immunol. 209, 46(16):3292-9. Thuslocalization and sequestration, possible in a limited sample butdifficult systemically, has been show to protect and improvedifferentiation in dendritic cells.

The invention of this application offers a unique opportunity to combineand maximize the effectiveness of two therapies otherwise limited by theex vivo nature of the treatment. By employing different modules 3008 a,3008 b, 3008 c, etc., in any sequence in this invention, the patient maybe treated by reduction of significant amounts of circulating Gal-3while being treated by, e.g., photopheresis—the resulting dendritic-celluptake and immune cell activation less likely to be defeated by thereduced level of circulating Gal-3. Thus, Gal-3 removal might beeffected in modules 3008 a and 3008 b, while photopheresis is practicedin module 3000 c. It is noted that Gal-3 removal can also take place asan independent procedure, preferably in close proximity time wise tophotopheresis, and preferably before photopheresis. While it isdifficult to measure the presence of synergy in an ex vivo treatmentregime, there is clearly an opportunity to enhance the effectiveness ofboth the reduction of inflammation by Gal-3 removal and the activationof immune cells, particularly through photopheresis, while minimizingthe tendency of Gal-3 to undermine the effectiveness of thosetreatments. One of the greatest problems encountered in many otherwiseeffective treatments is patient compliance. By combining thesetreatments in a multi-module process such as the one disclosed herein,multiple regimens are effected in the same time with efficiency,avoiding multiple patient sessions.

To further enhance these and other therapies, the modules employed inthe apheresis system of this invention need not be limited to removaland treatment actions. VEGF and TGF-ß for example are two well-studiedinhibitors of mammalian immune function. Ohm et al, Immunol. Res. 2001,23(2-3):263-72; Viel et al, Sol. Signal, 2016, 9(415). The art suggeststhat by addition of signaling receptor blockers, or by adding decoyreceptors, the signaling of TGF-ß may be limited, reducing the tendencyof this agent to promote tumor growth and cancer metastasis. Thus,removal of VEGF, TGF-ß and similar CIPFs constitute one aspect of thedisclosed invention. By the same measure, their receptors and receptorsfor Tumor Necrosis Factor (TNF-alpha and beta, primarily) have becometargets for therapeutic treatment, the removal or blocking of which isadvantageously combined with selective withdrawal of Gal-3. Thesetechniques are advantageously provided in one sitting by Applicant'sinvention.

At the same time active Gal-3 is removed, soluble TNF receptors, bothR-1 and/or R-2 at different ratios based on the condition, are removed,through the same process, by running the plasma fluid over a bed ofbinding agents of TNF receptors. TNF can then directly target cancercells or other targets as an effective treatment. The reduction ofactive Gal-3 in both the circulation and the tissue level will allow TNFto exert its beneficial effects with a reduced amount of inflammationand fibrosis which limits its use. Wu et al, Arch. Dermatol. 20:1-7(2012). The effective removal of serum Gal-3 also enhances chemotherapy,particularly, but not exclusively, when combined with TNF receptorremoval. This can be further combined with the administration, throughthe same modules, of TNF receptor inhibitors and ligands, which furtherimprove, systemically, the ability of TNF to target tumors, preferablyaided by the removal of Gal-3. The removal of TNF receptors, andaddition of TNF receptor binders, might be effectively provided for inmodulus 3008 y and 3008 z.

As illustrated above, the modules that are combined into the apheresissystem may be adapted to address the particular challenges presented forany particular patient. The ability to “print” columns provided with thenecessary functionality coupled with the ease with which functionalagents may be added to the same withdrawn blood or separated plasmapermit adaptation of the methods and system of this invention to anygiven patient's needs. As noted, in a single patient visit, targets suchas a galectin, including Gal-3, TNF alpha and beta receptors, cytokinesof various character may all be removed. In some patients, excessive TNFmay be removed by providing a column with specialized receptors. At thesame time, it may be possible to add agents whose effectiveness isenhanced by removal of inhibiting agents. Thus, where other CIPF's,inflammatory compounds, growth factors, etc. are removed, it may beadvantageous in the same treatment to add agents, such as variouspharmaceuticals and similar therapeutic agents, like chemotherapeuticagents, hormones, CAR-T Cells, etc. As noted above, the inventionembraces the sensitization of various cells, in a variety of modes suchas activated immune cells. Both host and donor cells may be involved.Those of skill in the art—medical doctors with familiarity with thecascade of problems presented by a single or multiple health issue, willselect those treatments best suited to address a patient's needs in asingle, comfortable setting. Improving patient compliance is just oneaspect of the improvements obtained by this invention.

This invention has been disclosed in terms of embodiments andalternatives readily identifiable to the inventor today. It goes withoutsaying that the pace of medical advance is such that tomorrow, newtargets for removal will be identified, new agents for addition will beadvanced, and new technologies for the design and manufacture ofpersonalized option will become manifest. The essential character ofthis invention is rather than relying on apheresis for the high costtreatment of a few diseases or syndromes, it may be used to tailorsuperior solutions on a patient-by-patient basis without introducingextraordinary costs or delays of heroic proportions in terms ofapprovals. By providing an apheresis device that may be personalized byswitching in or out a column or cartridge, custom made or specificallyprepared for the patient's needs, recognizing that in many if not mostcases those needs persist over time, the fundamental cost of apheresiscan be defrayed and its advantages employed in the treatment of awide-variety of conditions not previously adequately addressed byconventional options, and a more effective therapy can be offered.

1.-8. (canceled)
 9. A method for treating a mammal suffering from atumorous cancer in an ex vivo system, comprising: withdrawing an amountof blood from said mammal and introducing it, or plasma separatedtherefrom, into an ex vivo treatment system and subsequently returningsaid withdrawn blood to said mammal, following treatment, to saidmammal, wherein said method comprises: selectively withdrawing at leastten percent of galectin-3 in said withdrawn blood or plasma separatedtherefrom by passing said blood or plasma through a module provided witha moiety which selectively binds galectin-3, wherein said moiety isselected from the group consisting of an antibody, antibody fragment,non-antibody peptides, each of which selectively binds galectin-3, apolysaccharide which selectively binds galectin-3, or a combinationthereof; wherein said method further comprises administering to saidmammal at least one of a PD-1 inhibitor and a PD-L1 in conjunction withsaid withdrawal of blood.
 10. The method of claim 9, wherein said methodfurther comprises administering a dose of CAR-T cells to said patient inconjunction with said withdrawal of blood.
 11. The method of claim 9,wherein said method comprises treating said mammal with photodynamictherapy comprising exposure of said mammal to light selected so as toactivate a photosensitizer administered to said mammal.
 12. The methodof claim 9, wherein said method further comprises selectivelywithdrawing at least one of TNFα and TNFβ receptors from blood of saidmammal by passing said withdrawn blood through a module with a moietywhich selectively binds at least one of receptors for TNFα or receptorsfor TNFβ.
 13. A method for treating a mammal suffering from a tumorouscancer in an ex vivo system, wherein said method comprises withdrawingan amount of blood from said mammal and introducing it or plasmaseparated therefrom into an ex vivo treatment system and subsequentlyreturning said withdrawn blood to said mammal, following treatment, tosaid mammal, wherein said method comprises: selectively withdrawing atleast ten percent of galectin-3 in said withdrawn blood by passing saidblood through a module provided with a moiety which selectively bindsgalectin-3, wherein said moiety is selected from the group consisting ofan antibody, antibody fragment, non-antibody peptides, each of whichselectively binds galectin-3, a polysaccharide which selectively bindsgalectin-3, or a combination thereof; and selectively withdrawing atleast one of TNFα or TNFβ receptors from said withdrawn blood passingsaid withdrawn blood through a module provided with a moiety whichselectively binds at least one of TNFα or TNFβ receptors; said methodfurther comprising administering to said patient an effective amount ofCART-cells, a dose of PD-1 or PD-L1 inhibitor, or a photosensitizer forphotodynamic therapy for said mammal.
 14. A method for treating a mammalin an ex vivo system, wherein said method comprises withdrawing anamount of blood from said mammal and introducing it into an ex vivotreatment system and subsequently returning said withdrawn blood to saidmammal, following treatment, to said mammal, wherein said methodcomprises: selectively withdrawing at least ten percent of galectin-3 insaid withdrawn blood by passing said blood or plasma separated therefromthrough a module provided with a moiety which selectively bindsgalectin-3, wherein said moiety is selected from the group consisting ofan antibody, antibody fragment, non-antibody peptides, each of whichselectively binds galectin-3, a polysaccharide which selectively bindsgalectin-3, or a combination thereof; and selectively withdrawing atleast one inflammatory cytokine from said withdrawn blood by passingsaid withdrawn blood through a module provided with a moiety whichselectively binds said at least one inflammatory cytokine, prior toreturn of said withdrawn blood to said patient.
 15. The method of claim14, wherein said at least one inflammatory cytokine is selected from thegroup consisting of CXCL-8, CCL2, CCL3, CCL4, CCL5, CCL11, CXCL10, a CCLcytokine, a CXL cytokine, CX3CL, and an XCL cytokine.
 16. A method ofimproving the immune response of a mammal in need thereof, comprising:withdrawing an amount of blood from said mammal and introducing it intoan ex vivo treatment system and subsequently returning said withdrawnblood to said mammal, following treatment, to said mammal, wherein saidmethod comprises: selectively withdrawing, through apheresis, aneffective amount of at least one of TNFα receptors or TNFβ receptorspresent in said withdrawn blood by passing said blood or plasmaseparated therefrom through a module provided with a moiety whichselectively binds said TNFα or TNFβ; and administering to said mammal,in conjunction with said apheresis, an effective amount of at least oneof PD-1 or PD-L1 inhibitor.